Development and Validation of a HPLC Method for Determination of Pefloxacin in Tablet and Human Plasma

Objective(s) Developing and validating a simple, efficient, reproducible and economic reversed phase high performance liquid chromatographic (RP-HPLC) method for the quantitative determination of pefloxacin in bulk material, tablets and in human plasma. Materials and Methods A shim-pack CLC-ODS column and a mobile phase constituting acetonitrile: ۰.۰۲۵ M phosphoric acid solution (۱۳:۸۷ v/v, pH ۲.۹ adjusted with KOH) were used. The flow rate was ۱ ml/min and the analyses performed using ultraviolet (UV) detector at a wavelength of ۲۷۵ nm using acetaminophen as an internal standard. Results The developed method showed good resolution between pefloxacin and acetaminophen. It was selective to pefloxacin and able to resolve the drug peak from internal standard and from formulation excipients. The percentage of coefficient variation (CV) of the retention times and peak areas of pefloxacin from the six consecutive injections were ۰.۵۶۶% and ۰.۹۸۹%, respectively. The results showed that the peak area responses are linear within the concentration range of ۰.۱۲۵ qg/ml-۱۲ qg/ml (R۲= ۰.۹۹۸۷). The limits of detection (LOD) and limits of quantitation (LOQ) for pefloxacin were ۰.۰۳۱۲۵ qg/ml and ۰.۱۲۵ qg/ml. The intra-day and inter-day variation, RSD were ۰.۳۷۶-۰.۹۰۵۶ and ۰.۷۳۹-۰.۸۵۳ respectively; also, inter-day variation with relative standard deviation (RSD) were ۰.۱۴۶۵-۰.۸۲۱ in plasma. The accuracy results of ۷۰%, ۱۰۰%, and ۱۳۰% drugs were ۱۰۰.۷۲%, ۱۰۰.۳۴%, and ۱۰۰.۰۹%, respectively. Conclusion The method is linear, quantitative, reproducible and could be used as a more convenient, efficient and economical method for the trace analysis of drug in biological fluids, in raw material and tablets.

کلیدواژه ها:

Antibiotics ، fluoroquinolone ، High Performance Liquid Chromatography ، Pefloxacin ، Quantitative analysis ، Validation studies